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. 2023 Feb 14;14:820. doi: 10.1038/s41467-023-36445-9

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a U2OS WT and PKR KO cells were transfected with poly(I:C) (200 ng/mL, 16 h), and the number of G3BP1 foci by cell was quantified. Top; percentage of cells with G3BP1 foci. Red lines indicate the mean (Number of cells, n = 250). P-values were calculated with a two-tailed Welch t-test. b Quantification of G3BP1 foci area after transfection with poly(I:C) (200 ng/mL, 16 h) in U2OS-A3B-Flag WT and PKR-A3B-Flag KO cells. Red lines indicate the mean (Number of G3BP1 foci, n = 250). P-values were calculated with a two-tailed Welch t-test. c. Formation of G3BP1 foci after poly(I:C) (200 ng/mL, 16 h) transfection was monitored by immunofluorescence staining in U2OS-A3B-Flag WT and RNase L KO cells without DOX treatment. Scale bar: 5 μm. d U2OS-A3B-Flag WT, RNase L KO, PKR/RNase L DKO cells were transfected with poly(I:C) (200 ng/mL, 16 h), and the number of G3BP1 foci in individual cells were quantified without DOX treatment. Red lines indicate the mean (Number of cells, n = 250). P-values were calculated with a two-tailed Welch t-test. Number of G3BP1 foci by cell was quantified in U2OS-A3B-Flag PKR KO cells without DOX treatment and knocked down with A3B (e) or PABPC1 (f) siRNAs for 40 h and transfected with poly(I:C) (200 ng/mL, 4 h). Red lines indicate the mean (Number of cells, n = 250). P-values were calculated with a two-tailed Welch t-test. g Localization of G3BP1 and FAM120A were monitored by immunofluorescence in U2OS-A3B-Flag cells ±DOX after transfection with poly(I:C) (200 ng/mL, 16 h). Scale bar: 5 μm. h Quantification of FAM120A intensity (arbitrary units) in G3BP1 foci from experiment described in (g). Red lines indicate the mean (Number of G3BP1 foci, n = 250). P-values were calculated with a two-tailed Welch t-test. i SG formation through an RNase L-dependent mechanism and a PKR-dependent mechanism during viral infection. A3B and PABPC1 protect SGs from RNase L-induced RNA cleavage.